Reduction inhibitory agent for active oxygen eliminating activity

ABSTRACT

A reduction inhibitory agent for active oxygen eliminating activity comprises trehalose as an active ingredient. This agent can be incorporated into plant edible products and/or plant antioxidants.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application is a divisional of Ser. No. 09/034,336, filed Mar. 4, 1998, the entire contents of which are hereby incorporated by reference.

FIELD OF THE INVENTION

[0002] The present invention relates to an agent which inhibits reduction of active oxygen-eliminating activity, uses of the same, and a method for inhibiting reduction of active oxygen eliminating activity. More particularly, the present invention relates to an agent for inhibiting reduction of active oxygen eliminating activity which comprises trehalose as an active ingredient, uses of the same as compositions which can be obtained by incorporating the inhibitory agent into edible plant substances, and/or plant antioxidants, and a method for inhibiting the reduction of active oxygen eliminating activity of plant substances.

BACKGROUND OF THE INVENTION

[0003] It is well known that edible plant substances such as vegetables, mushrooms, and layers are important for living bodies as sources of functional ingredients such as vitamins, minerals, and edible fibers, and are essential food materials for the growth of living bodies. Recently, interest has focused on the active oxygen eliminating activity of these edible plants with respect to maintenance and promotion of health, prevention of aging and geriatric disease, and inhibition of carcinogenesis. Mechanisms for the causes of aging of living bodies and related diseases thereof such as cancers, arteriosclerosis, liver cirrhosis, myocardial infarction, cerebral apoplexy, cataracts, Parkinson's disease, rheumatism, and Alzheimer's dementia are still unknown in many aspects. However, there is some consideration that these incurable diseases may be related to active oxygen molecules such as superoxide, which is an oxygen molecule having an unpaired electron and relatively high reactivity, and derivatives thereof, including hydroxyradical and hydrogen peroxide. It is believed that these molecules oxidize intracellular target molecules such as membrane lipids, proteins, and DNAs to induce oxygen-related defects and cause aging of living bodies and related diseases thereof.

[0004] Enzymes capable of eliminating active oxygen such as superoxide dismutase (EC 1.15.1.1) and catalase (EC 1.11.1.6), and antioxidants such as L-ascorbic acid and α-tocopherol, exist in living cells, so that the concentration of intracellular active oxygen is kept generally at an extremely low level. However, irradiation from a relatively large amount of ultraviolet rays, radiations, and magnetic waves, excessive physical exercise, great mental stress, and aging form active oxygen in an amount that cannot be eliminated solely the active oxygen eliminating of living bodies. This results in an accumulation of compounds oxidized by the excessive amount of active oxygen which causes the aforesaid oxygen-related defects.

[0005] To improve the problem, for example, Japanese Patent Kokai Nos. 168,435/93 and 143,466/96 proposed methods which comprise supplementing living bodies with active oxygen eliminating activity to promote the maintenance and promoting of health of living bodies by using a relatively high active oxygen eliminating activity of edible plants. It was confirmed that even if such edible plants were used, processing techniques such as juice extraction, extraction, heating, and drying, as well as successive storage conditions for the plants, may lower the inherent active oxygen eliminating activity or even extinguish this activity completely. There is a need for establishing a novel method for inhibiting the reductions of active oxygen eliminating activity without causing serious side effects in living bodies.

SUMMARY OF THE INVENTION

[0006] The object of the present invention is to provide a novel agent for reducing active oxygen eliminating activity, a composition which can be used by incorporating the agent into edible plant substances and/or plant antioxidants to inhibit their active oxygen eliminating activity, and a method for inhibiting the reduction of active oxygen eliminating activity of edible plant substances and/or plant antioxidants.

[0007] To attain the above objects, the present inventors studied the use of saccharides, and continued studying. They examined the influence of reduction of the inhibitory effect on active oxygen eliminating activity of plant substances with active oxygen eliminating activity such as edible plant substances and plant antioxidants by incorporating saccharides therein.

[0008] As a result, the present inventors found that trehalose exerts a more effective activity than other saccharides and strongly inhibits reduction of plant active oxygen eliminating activity, and thus accomplished the present invention.

[0009] The first object of the present invention is to provide a reduction inhibitory agent for active oxygen eliminating activity which comprises trehalose as an effective ingredient. The second object of the present invention is to provide a composition which can be obtained by incorporating the agent into edible plant substances and/or plant antioxidants with active oxygen eliminating activity to inhibit the reduction of their active oxygen eliminating activity. The third object of the present invention is to provide a method for inhibiting the reduction of plant active oxygen eliminating activity which comprises incorporating the agent into plant substances with active oxygen eliminating activity.

DETAILED DESCRIPTION OF THE INVENTION

[0010] The trehalose suitable used as an effective ingredient in the present reduction inhibitory agent for active oxygen eliminating activity is α,α-trehalose, although any trehalose can be used in the present invention independent of source or origin, as long as it inhibits the reduction of active oxygen eliminating activity. For example trehaloses prepared from yeasts as disclosed in Japanese Patent Kokai No. 246,097/95, from maltose as disclosed in Japanese Patent Kokai Nos. 216,695/83 and 170,977/95, and from starch as disclosed in Japanese Patent Kokai Nos 143,876/95 and 213,283/95, can be used in the present invention. Any form of trehalose, such as syrup, crystalline powder of massecuite, crystalline hydrate, crystalline anhydride, or amorphous solid can be used. Commercially available high purity crystalline trehalose hydrate and crystalline trehalose anhydride can be arbitrarily used. The trehalose content in the present reduction inhibitory agent for active oxygen eliminating activity can be selected from those which exert a desired reduction inhibitory activity on the active oxygen eliminating activity of edible plant substance, usually, at least about 20 w/w % on a dry solid basis (d.s.b.), and preferably at least 50%, d.s.b. For purposes of the present specification, % means w/w % unless otherwise specified. The reduction inhibitory agent can be suitably incorporated into compositions containing edible plant substances and/or plant antioxidants in an amount of at least one percent, preferably at least 5 percent, and more preferably at least 20 percent, d.s.b.

[0011] The word “incorporating” in the present invention means “coexisting.” In the case of incorporating the present reduction inhibitory agent into plant substances with active oxygen eliminating activity, any conventional methods can be used as long as they allow the agent to impart satisfactory reduction inhibitory activity for plant active oxygen eliminating activity independent of the incorporation methods and relative proportions. The present agent can be preferably incorporated into plant edible substances and/or plant antioxidants by allowing the agent to contact the substances and/or the antioxidants in aqueous systems as homogeneously as possible.

[0012] For example, in the case of using edible plant substances, etc., in a juicy form such as a liquid or suspension form, the present agent in the form of a powder, crystal, or solid can be incorporated into the plant edible substances as homogeneously as possible by mixing with and dissolving in the substances. While in the case of using edible plant substances and plant antioxidants in a solid form, they are treated with water to give a juicy form such as a liquid or suspension form, and are incorporated with the present agent by the above method or providing a syrup of the present agent and dispersing, dissolving, or suspending the edible plant substances and antioxidants in the syrup to allow contact with the agent as homogeneously as possible to incorporate the agent into the substances and antioxidants. In the case of using edible plant substances such as disrupted raw plant tissues, the present agent can be incorporated into the edible plant substances either by sprinkling the agent over the substances and mixing them to dissolve the present agent in the substances, or by soaking the edible plant substances in the agent in a syrup form.

[0013] The plant substances with active oxygen eliminating activity which can be used in the present invention include edible plants per se; disrupted, minced, pulverized, dried, and extracted products of edible parts of the edible plants; and/or plant antioxidants. Examples of the edible plants suitably used in the present invention are edible root crops such as carrots, lotus roots, onions, burdocks, Japanese radishes, taros, and yams; leafy vegetables such as lettuces, chicories, Chinese cabbages, cabbages, kales, moreheiya or Corchorus olitorius, Angelica keiskei, spinach, Malabar nightshades, komatsuna (a kind of Chinese cabbage), nozawana (a kind of turnip), Chrysanthemum coronirium, chingensai (a Chinese vegetable), and turnips; fruiting vegetables such as okura or gumbos, cauliflowers, broccolis, eggplants, tomatoes, cucumbers, pumpkins, zucchini, green peppers, field peas, and kidney beans; vegetables such as alfalfa, bean sprouts including those of soy beans and mung beans; mushrooms such as shiitake mushrooms, winter mushrooms or Celtis sinensis, and shimeju mushrooms; seaweeds such as hijiki seaweed of Hijikia fusiformis, wakame seaweed or undaria pinnatifida, and tangs; citrus fruits such as lemons, citrons, and sudachi (a kind of citrus fruit), shaddocks, and kumquats; fruits such as bananas, kiwifruits, strawberries, hawthorns, blueberries, grapes, peaches, apples, pears, chestnuts; herbs such as garlic, gingers, wasabis (a Japanese horseradish), mustards, parsleys, Chinese parsleys, beefsteak plants, leeks, Welsh onions, celeries, dropworts, cressons, Guinea peppers, Japanese peppers, peppers, rosemaries, and mints; medicinal herbs and crude drugs such as mugworts, common plantains, bad-smelling perennial plants of the family Sauruaceae, Cassia obtusifolia, Japanese green gentians, aloes, licorices, turmerics, Japanese indigo plants, Pfaffia, loquat leaves, field horsetails, pine needles, bamboo leaves, umes or Japanese apricots, tea leaves, barley leaves, wheat leaves, oat leaves, rye leaves, buckwheat leaves, ginkgo leaves, Chinese gutta percha leaves, oobanasarusuberi (a plant of the family Lythrum), Aspalathus linearis, and Gymnema sylvestre; and others including edible parts of nuts, seed, and germs such as adlays, buckwheat's, sesames, rices, wheats, corns, broad beans, soybeans, peanuts, walnuts, pine seeds, and other seeds and germs.

[0014] One or more of the following relatively low molecular weight plant antioxidants can be suitably selected and used as the present plant antioxidants: plant enzymes having active oxygen eliminating activity such as superoxide dismutase, catalase, and superoxide; plant pigments such as rutin, α-glycosyl rutin, hesperidin, α-glucosyl hesperidin, naringin, α-glucosyl naringin, chlorophyll, carotenoid, and anthocyanin; plant polyphenols such as gallic acid, catechin, α-glucosyl catechin, and tannic acid; and plant vitamins such as α-tocopherol, L-ascorbic acid, and riboflavin. In the present invention, the relatively low molecular weight plant antioxidants can be contained in the edible plants or added to the contents during processing of the edible parts of edible plants. The plant antioxidants, which exist separately from the edible plants, can be used in the present invention. Plant antioxidants should not be restricted to those which are extracted from plants, and include those which are artificially synthesized and produced.

[0015] More concretely, processes for producing compositions according to the present invention, in which the reduction of the active oxygen eliminating activity is satisfactorily inhibited, as are follows:

[0016] Edible parts of edible plants are, for example,

[0017] (i) disrupted or cut by mixers, juicers, or pulverizers into pastes or suspensions, into which the present agent is incorporated by adding it thereto and dissolving it therein; or

[0018] (ii) extracted with solvents such as hot water and alcohol and filtered, if necessary, further concentrated into liquids or pastes, followed by incorporating the present agent into the liquids or pastes by mixing to dissolve it therein.

[0019] The compositions thus obtained are usually syrupy, pasty, or juicy disrupted products which have a relatively high moisture content and require a relatively low storage temperature for a satisfactory shelf life. The compositions can be dried and/or pulverized, or dried and/or pulverized after sterilization by heating to obtain powdery or small pieces of solid compositions. Conventional drying and pulverizing methods can be arbitrarily used in the present invention. The compositions of syrupy, pasty, or juicy disrupted products can be

[0020] (i) successively dried in vacuo, air dried, drum dried, pulverized, and sieved;

[0021] (ii) successively spray dried, subjected to fluidized bed drying, and sieved; or

[0022] (iii) successively mixed with anhydrous saccharides such as maltose or trehalose anhydride to effect dehydration, dried, and sieved into powdery products.

[0023] If necessary, the powdery products can be arbitrarily granulated, tabletted, and encapsulated into appropriate shapes and forms for use. The solid compositions, produces to such a high degree, will acquire more improved activity for inhibiting the reduction of active oxygen eliminating activity, storage stability, and handelability. The solid compositions containing edible plant substances have features such that they effectively inhibit the deterioration of plant pigments contained in the edible plants, such as chlorophylls, carotenoids, anthocyanins, and flavonoids. Additionally, they satisfactorily retain the inherent color of the pigments and mask undesirable tastes such as bitterness and astringency.

[0024] Compositions according to the present invention containing trehalose and a plant edible substance or plant antioxidant and which inhibit reduction of active oxygen eliminating activity can be further mixed with one or more appropriate substances in the form of a solid or liquid, such as essential minerals, edible fibers saccharides for promoting the growth of bifid bacteria, vitamins, biologically active substances, and preservatives.

[0025] Essential minerals which can be used in the present invention include, for example, calcium, magnesium, phosphorus, iron, copper, zinc, and cobalt. These minerals are used in an adequate amount.

[0026] Edible fibers which can be used in the present invention are, for example, pectin, alginic acid, carrageenan, gum Arabic, glucomannan, cyclodextrin, and pullulan. Of the edible fibers, it has been discovered that pullulan in particular has a strong reduction inhibitory activity for —plant active oxygen eliminating activity when used in combination with trehalose as the effective ingredient according to the present invention. Based on this, pullulan can be most satisfactorily used together with trehalose in the present agent and used to produce compositions according to the present invention.

[0027] Examples of saccharides which can be used to promote the growth of bifid bacteria according to the present invention are lactosucrose, fructooligosaccharide, galactooligosaccharide, and isomaltooligosaccharide, which can arbitrarily used on demand.

[0028] Vitamins which can be used in compositions according to the present invention include, for example, water soluble vitamins such as thiamin, riboflavin, L-ascorbic acid, rutin, hesperidin, naringin, niacin, pyridoxine, cyanocobalamin, and derivatives thereof. Lipid-soluble vitamins which can be used include vitamin A, vitamin D, α-tocopherol, vitamin K, and derivatives thereof.

[0029] One or more biologically active substances can be incorporated into the compositions of the present invention, including hormones, antibiotics, cytokines, and propolis. Adequate amounts of ethanol, acetic acid, lactic acid and salt can optionally be used as preservatives in compositions according to the present invention.

[0030] To improve the quality of the compositions of the present invention as well as other compositions containing these compositions, adequate coloring agents, flavoring agents, taste-imparting agents, stabilizers, fillers, adjuvants, and excipients can be used.

[0031] The compositions thus obtained have at least five units/g composition, d.s.b., and preferably at least 10 units/g, d.s.b., of active oxygen eliminating activity as determined by the nitroblue tetrazolium (NBT) test. These compositions can easily administered to subjects via appropriate routes, including orally, intubationally, percutaneously, and permucosally to impart active oxygen eliminating activity with lesser deterioration of their quality. Intake of the composition of the present invention containing plant edible substances enriches plant functional ingredients such as vitamins, mineral, and edible fibers inherent in the plants, and enriches living bodies with active oxygen eliminating activity. Thus, compositions of the present invention can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and inhibit carcinogenesis. The compositions of the present invention can be used in food products, cosmetics, pharmaceuticals, and their components and processing intermediates.

[0032] To enrich nutritional value and improve qualities and taste, composition of the present invention can be used for health food products and other food products in general. Examples of these are seasonings such as “furikaka” (a seasoned fish meal), sauces, ketchup, “yakinikui-no-tare” (a sauce for Japanese grilled meat). Curry roux, and instant soup mixes; Japanese and Western cakes such as “mochi” (a rice paste), “dango” (a ball of rice paste), candies, chewing gum, baked confections, snack confections, waffles, sponge cakes, buns, and baked goods; frozen desserts such as ice cream, sherbet, and candies; pastes such as fruit pastes, fruit sources, peanut pastes, and raw jams; pickles and pickled products; meat products such as hams and sausages; products of fish meats such as “kaaboko” (a steamed fish paste), “chikuwa” (a kind of fish paste), “hanpen” (a fish cake), and sausages; “chinmi” (a delicacy); “tsukudani” (a food boiled down in soy); dairy dishes; beverages such as vegetable juices, fruit juices, and carbonated beverages; rice products such as rice gruel, porridge of rice and vegetables, and cooked rice with seasonings, meats, and vegetables; noodles such as iced noodles, macaroni, and pastes; food products such as pudding and cake mixes, instant juices, instant soups, and frozen foods; as well as their components and processing intermediates.

[0033] Compositions according to the present invention can be used in cosmetics such as skin-beautifying agents, packs, creams, shampoos, hair rinses, hair tonics, bath salts, enzymic agents, and tooth pastes in the form of a liquid, paste, powder, or granule; and used as a component of a processing intermediate for these products.

[0034] Compositions according to the present invention can be used to treat age-related diseases in the form of an oral and/or intubation nutrient or therapeutic agent such as a coilunarium, nebula, digestive agent, stomachic, enzymic agent, ointment, cataplasm, and their components and processing intermediates.

[0035] Compositions of the present invention comprises substantial food materials and can be used with less fear of causing toxicity. Usually the compositions can be used on a dry solid basis at a dose of about 0.1 to 1000 g/adult/day, preferably, about 0.2 to 500 g/adult/day, and more preferably, about 1 to 11 g/adult/day when administered orally.

[0036] Compositions of the present invention can be administered to subjects intubationally, percutaneously, and permucosally in accordance with the oral dosage.

[0037] The following experiments describe the present invention in detail:

[0038] Experiment 1: Influence of Saccharides on Reduction Inhibitory Activity for Active-oxygen Eliminating Activity

[0039] Fresh raw carrots were disrupted by a mixer, and 10% of either of saccharides, d.s.b., was added to the mixture and dissolved therein. The solution was dried under reduced pressures at 40° C. for two days, dried in vacuo at the same temperature for 24 hours, and pulverized by a mixer into a powdery carrot composition. The saccharides used were commercialized reagents of glucose, mannitol, sorbitol, maltose, sucrose, trehalose, and pullulan. An about 100 g of the composition was placed and sealed in a 500-ml polystyrene container, and stored at 40° C. for seven days. The active-oxygen eliminating activity of the composition before and after the storage were compared. The assay for the active-oxygen eliminating activity followed the nitroblue tetrazolium test by Toshio IMANARI et al. in Igakuno Ayumi, Vol. 101, pp. 496-497 (1977); the level of superoxide, formed in a xanthine-xanthine oxidase system, was assayed by quantifying calorimetrically the content of formazan formed by reducing NBT. As a control, refined water was used as a test solution. One unit activity of active-oxygen eliminating activity is defined as that inhibits 50% formazan formation as compared with the control. Table 1 shows the composition of the powdery compositions, and the active-oxygen eliminating activity per gram of either of the compositions before and after the storage. As evident from Table 1, among the saccharides tested the composition with trehalose showed the highest residual percentage for active-oxygen eliminating activity, revealing that trehalose exerts a strong effect on the reduction inhibitory activity for active-oxygen eliminating activity. The reduction inhibitory activity of trehalose was strongly higher than that of glucose as the constituent saccharide for trehalose, and that of maltose as a disaccharide consisting of two glucose units similar to trehalose. Thus trehalose is suitably used as an effective ingredient for a reduction inhibitory agent for active-oxygen eliminating activity. Next to trehalose, pullulan showed a relatively-high reduction inhibitory activity for active-oxygen eliminating activity. TABLE 1 Composition of powdery Saccharide Activity (unit/g) Percentage of composition (%) content Before After residual Saccharide Carrot Saccharide Moisture (d.s.b.) (%) Storage Storage activity (%) None 93.4 0.0 6.6 0.0 870 100 11 Glucose 48.8 46.4 4.8 48.7 580 170 29 Mannitol 49.8 47.5 2.7 48.8 520 59 11 Sorbitol 48.2 46.0 5.8 48.8 560 26 5 Maltose 48.1 45.8 6.1 48.8 550 180 33 Sucrose 48.2 46.0 5.8 48.8 660 180 27 Trehalose 47.9 45.7 6.4 48.8 580 380 66 Pullulan 47.7 45.5 6.8 48.8 710 360 51

[0040] Experiment 2. Influence of Trehalose Concentration of Reduction Inhibitory Activity for Active-oxygen Eliminating Activity

[0041] Similarly as in Experiment 1, 0-20% of trehalose, d.s.b., was mixed with and dissolved in a distrupted carrot. The resulting mixture was dried in vacuo at 40° C. for 24 hours, and pulverized into a powdery carrot composition. Similarly as in Experiment 1, each powdery composition was placed and sealed in a polystyrene container, and stored at 40° C. for seven days for evaluating the reduction inhibitory activity for active-oxygen activity. Table 2 shows the active-oxygen eliminating activity of one gram of each powders composition before and after the storage. Based on the data from Table 2, at least one percent, preferably, at least five percent, and more preferably, at least 20% of trehalose, d.s.b., exerted a desired effective reducing inhibitory activity for active-oxygen eliminating activity. TABLE 2 Trehalose Percentage (%) of Composition of content Activity (unit/g) Percentage of trehalose added powdery composition (%) (d.s.b.) Before After residual activity to carrot Carrot Trehalose Moisture (%) storage Storage (%) 0.0 95.3 0.0 4.7 0.0 940 330 35 0.1 94.3 1.0 4.7 1.0 930 400 43 0.5 90.4 4.7 4.9 4.9 750 460 61 1.0 85.9 8.9 5.2 9.4 860 480 56 2.5 75.3 19.6 5.1 20.7 700 460 66 5.0 62.3 32.4 5.3 34.2 600 370 62 10.0 46.1 48.1 5.8 51.1 400 330 83 20.0 30.3 63.1 6.6 67.6 380 240 63

[0042] Experiment 3. Influence of Trehalose on Reducing Inhibitory Activity for Active-oxygen Eliminating Activity of Vegetables

[0043] Vegetables were treated similarly as in Experiment 1, and 10% trehalose, d.s.b., to the disrupted vegetables. The resulting mixture was dried in vacuo at 45° C. for 20 hours, and pulverized into a powdery vegetable composition. Similarly as in Experiment 1, each composition was respectively placed in a container and stored at 40° C. for six days. The active-oxygen eliminating activity of one gram of each composition before and after the storage was compared with that of the composition prepared without trehalose. Table 3 shows the composition of the powdery compositions, and their active-oxygen eliminating activity. Reduction inhibitory activity for active-oxygen eliminating activity by incorporation of trehalose was observed in vegetables such as carrot, onion, Japanese radish, cabbage, spinach, cucumber, and pumpkin. TABLE 3 Activity Percentage Composition of powdery Trehalose (unit/g) of residual Vegetable composition (%) content (d.s.b.) Before After activity (moisture (%)) Carrot Saccharide Moisture (%) storage storage (%) Carrot 94.5 0.0 5.5 0.0 1200 370 31 (90.1) 46.2 46.6 7.2 50.2 570 460 81 Onion 93.2 0.0 6.8 0.0 440 210 48 (89.3) 47.5 44.3 8.2 48.3 300 230 77 Japanese radish 92.7 0.0 7.3 0.0 1100 630 57 (94.0) 35.0 58.3 6.7 62.5 530 450 85 Cabbage 94.8 0.0 5.2 0.0 4100 2500 63 (93.8) 35.7 57.5 6.8 61.7 1600 1400 88 Spinach 93.2 0.0 6.8 0.0 3500 1300 36 (94.1) 34.4 58.4 7.2 62.9 1200 890 74 Eggplant 91.5 0.0 8.5 0.0 38000 31000 82 (93.7) 34.8 55.3 9.9 61.4 13000 11000 85 Cucumber 92.7 0.0 7.3 0.0 4600 1300 30 (95.0) 30.3 60.5 9.2 66.6 1900 1100 58 Pumpkin 93.2 0.0 6.8 0.0 1600 610 38 (84.0) 56.2 35.2 8.6 38.5 880 460 52

[0044] Experiment 4. Reduction Inhibitory Activity for Active-oxygen Eliminating Activity by Trehalose with Respect to Superoxide Dismutase and Antioxidants

[0045] In one milliliter of a 45 w/v % trehalose solution was dissolved five micrograms of superoxide dismutase from a horseradish commercialized by Sigma Chemical Co., St. Louis, USA, 10 mg α-glucosyl hesperidin commercialized by Hayashibara Biochemical Laboratories, Inc., Okayama Japan, 10 mg α-glucosyl rutin commercialized by Hayashibara Biochemical Laboratories, Inc., Okayama, Japan, 250 μg gallic acid commercialized by Wako Pure Chemical Industries, Ltd., Tokyo, Japan, or 50 μg catechin to obtain a liquid composition. Each of the compositions was placed in a glass test tube, sealed with a rubber cap, and stored under light-shielded conditions. As a control, water was used in place of the trehalose solution. Table 4 shows the active-oxygen eliminating activity before and after the storage. As shown in Table 4, it was confirmed that trehalose exerts an effective reducing inhibitory activity for active-oxygen eliminating activity of plant antioxidants such as superoxide dismutase, α-glucosyl hesperidin, α-glucosyl rutin, gallic acid, and catechin. Mere mixing of a powdery crystalline trehalose hydrate and either of the powdery antioxidants into a powders mixture, almost no change of the active-oxygen eliminating activity was observed when subjected to storage test similarly as above. Based on the results, it was revealed that a reducing inhibitory agent for active-oxygen eliminating activity, which comprises trehalose as an effective ingredient, could scarcely exert the desired activity only when mixed in a powder form with powdery substances having active-oxygen eliminating activity, but exert the desired activity when trehalose in a melted state is allowed to contact with and incorporate into the substances, or when trehalose is incorporated into the substances in an aqueous system. TABLE 4 Activity (unit/mg antioxidant) Content of Storage Trehalose Before After Residual Activity antioxidant Concentration concentration (%) storage storage (%) Superoxide dismutase 60° C. 0 10,000 4,600 46 (5 μg/ml) 17 hours 45 11,000 9,400 85 α-Glucosyl hesperidin 80° C. 0 3.8 1.0 26 (10 mg/ml) 5 days 45 3.6 2.4 67 α-Glucosyl rutin 80° C. 0 25 18 72 (10 mg/ml) 5 days 45 26 22 85 Gallic acid 80° C. 0 590 390 66 (250 μ/ml) 5 days 45 570 470 82 Catechin 25° C. 0 470 2.2  0 (50 μ/ml) 20 hours 45 380 310 82

[0046] Experiment 5. Reduction Inhibitory Effect for Active-oxygen Eliminating Activity on Herb Extracts

[0047] Ten liters of water was added to 500 g of a dried specimen of plantain, Plantago asiatica, commercialized by Daido Seiyaku Co., Ltd., Tokyo, Japan, and the mixture was infused for 90 min, followed by collecting a supernatant of the infused solution by a basket-type centrifuge. The supernatant was boiled down to obtain two kilograms of about 5 w/v % solution. Two hundred and forty grams of trehalose and 10 g of pullulan, d.s.b., were mixed with and dissolved in one kilogram of the above concentrate, and the solution was spray dried by “SD-1”, a portable spray-dryer commercialized by Tokyo rika Mfg. Co., Ltd., Tokyo, Japan, into a composition containing an extract of Plantago asiatica. As a control, using dextrin with DE (dextrose equivalent) of about 11 in place of the trehalose and pullulan, a powdery composition similar to the above composition was prepared. Similarly as in Experiment 1, these powdery compositions were placed in polystyrene containers, sealed, and stored at 40° C. for seven days for examining the reduction inhibitory effect for active-oxygen eliminating activity. Both the compositions had an activity of about 5,000 units/g composition before the storage. On seven days after the storage, the composition with trehalose and pullulan had an activity of about 4,300 units/g composition, while the composition with dextrin had an activity of about 3,100 units/g composition. The data shows that the use of trehalose and pullulan exerts a higher reduction inhibitory effect for active-oxygen eliminating activity than in the case of using dextrin.

[0048] The following Examples described the present invention; Examples A described the present reducing inhibitory agent for active-oxygen eliminating activity, and Examples B described the present composition where the reduction of active-oxygen eliminating activity is satisfactorily inhibited:

EXAMPLE A-1 Reduction Inhibitory Agent for Active-oxygen Eliminating Activity

[0049] Corn starch was prepared into an about 33% starch suspension which was then mixed with 0.1% calcium carbonate, adjusted to pH 6.5, mixed with 0.3 per g starch of “TERMAMYL”, an α-amylase specimen commercialized by Novo Industri A/S Copenhagen, Denmark, enzymatically reacted at 95° C. for 15 min, autoclaved at 120° C., and promptly cooled to obtain a liquefied solution with DE of about four. Thereafter, the liquefied solution was admixed with four units/g starch, d.s.b., of respective non-reducing saccharide-forming enzyme and trehalose-releasing enzyme, prepared by culturing a microorganism of the genus Rhizobium by the method in Japanese Patent Kokai No. 213,283/93, 500 units/g starch of isomaylase, and five units/g starch of cyclomaltodextrin glucanotransferase, adjusted to pH 6.2, and enzymatically reacted at 40° C. for 48 hours. The reaction mixture was heated to inactivate the remaining enzymes, then admixed with 10 unites/g substrate of glucoamylase and enzymatically reacted at pH 5.0 and 50° C. for 10 hours. The reaction mixture thus obtained contained about 86% trehalose, d.s.b. The mixture was heated to inactivate the remaining enzyme, then in a conventional manner purified by decoloring and desalting, and continuously crystallized while concentrating. The resulting massecuite was separated by a basket-type centrifuge, and the crystal was washed by spraying with a small amount of water to obtain a crystalline trehalose hydrate with a purity of 98% or higher in a yield of about 64%, d.s.b. The product is a crystalline trehalose hydrate with a considerably-high purity and suitably used as the present reduction inhibitory agent and advantageously used to inhibit the reduction of active-oxygen eliminating activity of plant edible substances.

EXAMPLE A-2 Reduction Inhibitory Agent for Active-oxygen Eliminating Activity

[0050] A microorganism of the genus Thermus capable of producing a maltose/trehalose converting enzyme was cultured in a nutrient culture medium by the method in Japanese Patent Kokai No. 170,977/95. After completion of the culture, the proliferated cells were collected by centrifugation to obtain 250 g wet cells having a total activity of about 7,500 units. The cells were immobilized using sodium alginate solution and calcium chloride solution to obtain alginic acid-immobilized cells. A jacketed-glass column, 5.4 cm in diameter and 100 cm in length, injected with the immobilized cells, and kept at 60° C. A 40% maltose solution (pH 6.5) was fed to the column at SV (space velocity) 0.2 by the descending method to obtain a reaction mixture containing about 66% trehalose, about 28% maltose, and about 6% glucose. The reaction mixture was in a conventional manner purified by decoloring and desalting, concentrated in vacuo into an about 75% syrup which was then placed in crystallizer, admixed with about one percent of respective seed crystals of crystalline trehalose hydrate and crystalline maltose hydrate, and cooled to obtain a massecuite with a crystallization degree of about 25%. The massecuite was spray dried and aged to obtain a powdery crystalline saccharide in a yield of about 92% to the material maltose, d.s.b. The powdery saccharide is a stable powdery saccharide, which has a crystallization degree of about 60% and contains about 66% trehalose, about 28% maltose, and about six % glucose, and it can be suitably used as a reduction inhibitory agent for active-oxygen eliminating activity and arbitrarily used to inhibit the reduction of active-oxygen eliminating activity of plant edible substances.

EXAMPLE A-3 Reduction Inhibitory Agent for Active-oxygen Eliminating Activity

[0051] Potato starch was prepared into a 10% starch suspension which was then subjected to the action of α-amylase to obtain a liquefied solution. To the liquefied solution were added three units/g starch, d.s.b., or a non-reducing saccharide-forming enzyme disclosed in Japanese Patent Kokai No. 213,283/95, five units/g starch, d.s.b., of a trehalose-relaeasing enzyme, 1,000 units/g starch, d.s.b., of an isoamylase, and one unit/g starch, d.s.b., of a maltotetraose-forming amylase, and the mixture was enzymatically reacted at pH 6.0 and 40° C. for 48 hours. The reaction mixture was heated to inactivate the remaining enzyme, and in a conventional manner purified by decoloring and desalting and concentrated in vacuo to obtain a syrup with a moisture content of about 30% and DE of about 15 in a yield of about 90%, d.s.b. The product is a stable syrup, which contains, on a dry solid basis, about 50% trehalose along with other saccharides derived from starch, can be suitably used as a reduction inhibitory agent for active-oxygen eliminating activity, and arbitrarily used to inhibit the reduction of active-oxygen eliminating activity of plant edible substances.

EXAMPLE A-4 Reduction Inhibitory Agent for Active-oxygen Eliminating Activity

[0052] One hundred parts by weight of a high-purity crystalline trehalose hydrate, obtained by the method in Example A-1, was mixed to homogeneity with one part by weight of pullulan to produce a solid reduction inhibitory agent for active-oxygen eliminating activity. The product can be arbitrarily used in a reduction inhibitory agent for active-oxygen eliminating activity of plant edible substances.

EXAMPLE A-5 Reduction Inhibitory Agent for Active-oxygen Eliminating Activity

[0053] In 100 hundred parts by weight of a trehalose syrup, obtained by the method in Example A-3, was dissolved by mixing to homogeneity 0.5 part by weight of pullulan, and 0.5 part by weight of a readily-water-soluble cyclodextrin into a syrupy reduction inhibitory agent for active-oxygen eliminating activity. The product can be arbitrarily used in a reduction inhibitory agent for active-oxygen eliminating activity of plant edible substances.

EXAMPLE B-1 Composition Containing Spinach

[0054] Fresh spinach was pulverized by a mixer, and in 10 parts by weight of the resulting disruptant were dissolved by mixing one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-4, and 0.1 part by weight of α-glucosyl rutin. The resulting mixture was heated at 100° C. for 10 min, dried with air heated to 40° C. for two hours, dried in vacuo at 40° C. for 16 hours, and powdered by a pulverizer to obtain a composition containing spinach having an active-oxygen eliminating activity of about 1,200 units/g composition.

[0055] The composition retained satisfactorily the inherent color of spinach and stimulated the appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the spinach, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-2 Composition Containing Cabbage

[0056] Cabbage was cut, branched, sliced by a cutter into fine strips about 5 mm wide. Ten parts by weight of the fine strips was mixed with one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-3, and 0.05 part by weight of α-glucosyl rutin, and the mixture was allowed to stand at ambient temperature for two hours, then dried by air heated to 40° C. for two hours, dried in vacuo at 40° C. for 16 hours to obtain a composition containing fine strips of cabbage having an active-oxygen eliminating activity of about 1,300 units/g composition. The composition retained satisfactorily the inherent color of the cabbage and stimulated your appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, and edible fibers inherent to the cabbage, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-3 Composition Containing Eggplant

[0057] A fresh eggplant was sliced, and the sliced eggplant was washed with water. One part by weight of a reduction inhibitory agent for active-oxygen eliminating activity, obtained by the method in Example A-1, was mixed with and dissolved in 10 parts by weight of the sliced eggplant, and the resulting mixture was dried by air heated to 70° C. for two days to obtain a composition containing sliced eggplant having an active-oxygen eliminating activity of about 11,000 units/g composition. The composition retained the inherent color of the eggplant's skin and stimulated the appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the eggplant, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-4 Composition Containing Carrot

[0058] A fresh carrot was branched and sliced by a slicer. One part by weight of a reduction inhibitory agent for active-oxygen eliminating activity, obtained by the method in Example A-4, was added to 10 parts by weight of the sliced carrot and dissolved therein. The mixture was dried with air heated to 50° C. for 16 hours to obtain a composition containing sliced carrots having an active-oxygen eliminating activity of about 450 units/g composition. The composition satisfactorily retained the color inherent to the carrot, stimulated the appetite, and had a relatively-low hygroscopicity and a satisfactory shelf-life. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the carrot, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-5 Composition Containing Hizikia fusiforme

[0059] Dried Hizikia fusiforme was rehydrated in water and disrupted with a cutter. In 10 parts by weight of the disrupted mixture was dissolved by mixing five parts by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-2. The solution was dried by air heated to 50° C. for eight hours, dried in vacuo at 40° C., and pulverized into a composition containing Hizikia fusiforme having an active-oxygen eliminating activity of about 24 units/g composition. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to Hizikia fusiforme, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-6 Composition Containing Shiitake Mushroom

[0060] A dried shiitake mushroom was rehydrated in water, heated at 100° C. for 15 min, and disrupted by a cutter. In 100 parts by weight of the disrupted mixture was dissolved by mixing 0.1 part by weight of a tea extract and 10 parts by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method in Example A-3, and the resulting mixture was dried at 50° C. for eight hours, dries in vacuo at 40° C., and pulverized to obtain a composition containing a powdery shiitake mushroom having an active-oxygen eliminating activity of about 560 units/g composition. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to Hizikia fusiforme, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-7 Composition Containing Citron

[0061] Citron rind was disrupted by a cutter, and five parts by weight of the disrupted rind was mixed with one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity, obtained by the method in Example A-1, to dissolve the agent therein. After preliminary freezing, the resulting mixture was freeze-dried for three days, and disrupted to obtain a composition containing a powdery citron having an active-oxygen eliminating activity of about 12 units/g composition. The composition retains satisfactorily the color and flavor inherent to the citron. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the citron, and imparts the active-oxygen eliminating activity to living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-8 Composition Containing Ginger

[0062] A fresh ginger was disrupted by a mixer. In five parts by weight of the disrupted mixture was dissolved by mixing one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-2. After preliminary freezing, the resulting mixture was freeze-dried for three days, and disrupted by a crusher to obtain a composition containing a powdery ginger having an active-oxygen eliminating activity of about 120 units/g composition. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the ginger, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote your health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-9 Composition Containing Aojiso (a Beefsteak Plant)

[0063] Aojiso leaves were branched and disrupted by a mixer. In five parts by weight of the disrupted mixture was dissolved by mixing one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-4, and 0.5 part by weight of α-glucosyl rutin, then mixed with 49 parts by weight of crystalline trehalose anhydride as a desiccant. The resulting mixture was allowed to stand at ambient temperature for one day and disrupted by a crusher to obtain a composition containing a powdery aojiso having an oxygen eliminating activity of about 230 units/g composition. The composition satisfactorily retained the color, flavor and taste of the aojiso, and stimulated the appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the aojjiso, and imparts the active-oxygen eliminating activity to living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE 10 Composition Containing Mugwort

[0064] According to a conventional manner, mugwort was placed into boiling water with an adequate amount of salt to remove harshness, dehydrated softly and disrupted by a crusher. Five parts by weight of the disrupted mixture was mixed with 0.05 part by weight of α-glucosyl rutin and 1.3 parts by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-5. The resulting mixture was dried by air heated to 50° C. for four hours, dried in vacuo at 40° C. for 16 hours, and disrupted to obtain a composition containing a powdery mugwort having an active-oxygen eliminating activity of about 780 units/g composition. The composition satisfactorily retained the color, flavor and taste of the mugwort, and stimulated the appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to mugwort, and imparts the active-oxygen eliminating activity to the living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-11 Composition Containing Dokudami (a Bad-smelling Perennial Plant of the Family Saururaceae

[0065] A half part by weight of a dried dokudami was mixed with 10 parts by weight of water, and heated to boiling for 90 min. A supernatant of the infused solution was collected by a basket-type centrifuge and boiled down to obtain four parts by weight of about five w/v % solution. Two parts by weight of he concentrate was mixed with 0.5 part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-4, and the resulting solution was spray-dried in a conventional manner to obtain a composition containing a powdery dokudami extract having an active-oxygen eliminating activity of about 1,400 units/g composition. The composition had a lesser unsatisfactory smell and taste inherent to the dokudami and could be more easily swallowable than conventional ones. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, an edible fibers inherent to the dokudami, and imparts the active-oxygen eliminating activity to living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-12 Composition Containing Aloe

[0066] A fresh aloe was disrupted by a mixer, and 10 parts by weight of the disrupted aloe was mixed with 0.1 part by weight of a tea extract and one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-1. The resulting mixture was dried by air, heated to 70° C. for two days, and disrupted by a crusher to obtain a composition containing a powdery aloe having an active-oxygen eliminating activity of 5,800 units/g composition. The composition had a lesser astringency of the aloe and was more easily swallowable than conventional ones. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the aloe, and imparts the active-oxygen eliminating activity to living bodies. The composition can be satisfactorily used to maintain and promote health, prevent aging and geriatric diseases, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus the composition can be arbitrarily used as a food product, cosmetic, pharmaceutical, or their material or processing intermediates.

EXAMPLE B-13 Composition Containing Chinese Cabbage Preserved with Seasonings

[0067] A Chinese cabbage preserved with seasonings was cut. To 10 parts by weight of the cut product was added and dissolved therein one part by weight of a reduction inhibitory agent for active-oxygen eliminating activity obtained by the method in Example A-1, and the mixture was dried with air heated to 55° C. for 16 hours to obtain a composition containing small pieces of Chinese cabbage, preserved with seasonings, having an active-oxygen eliminating activity of about 5,400 units/g composition. The product retained the color tint of the Chinese cabbage. When tasted, the satisfactory taste and flavor spread throughout the mouth and stimulated the appetite. Intake of this composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to the Chinese cabbage, and augments the active oxygen eliminating activity to the living bodies. The compositions can be satisfactorily used to maintain and promote health, prevent aging and geriatric disease, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or a component of processing intermediate thereof. The compositions can also be used as food products such as furikake, premix for onigiri (a rice ball), and soup for Chinese noodles, and as a health food, cosmetic, pharmaceutical, or components and intermediates thereof.

EXAMPLE B-14 Composition of Nozawana (a Kind of Turnip)

[0068] A pickled nozawana was subjected to a cutter. To 10 parts by weight of the cut product was added and dissolved therein one part by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method in Example A-1. The mixture was dried in air, heated to 55° C. for 16 hours to obtain a composition containing small pieces of pickled nozawana having an active oxygen eliminating activity of about 2600 units/g composition. The product retained the color tint of the picked nozawana. When tasted, the satisfactory taste and flavor spread throughout the mouth and stimulated the appetite. Intake of this composition enriches living bodies with functional ingredients such as vitamins, minerals and edible fibers inherent to the pickled nozawana, and augments the active oxygen eliminating activity to the living bodies. The composition can be used to maintain and promote health, prevent aging and geriatric diseases, promote treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-15 Composition of Korean Pickled Chinese Cabbage

[0069] Korean pickled Chinese cabbage was cut. To ten parts by weight of the cut product was added and dissolved therein was added one part by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method of Example A-4. The mixture was air dried and heated to 55° C. for 16 hours to produce a composition containing small pieces of pickled Chinese cabbage having an active oxygen eliminating activity of about 1600 units/g composition. The product retained the color tine of Korean pickled Chinese cabbage. When tasted, the satisfactory taste and flavor spread throughout the mouth and stimulated the appetite. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers inherent to Korean picked Chinese cabbage, and augments the active oxygen eliminating activity to the living bodies. The composition can be used to maintain and promote health, prevent aging and geriatric disease, promotes the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-16 Composition Containing Soybeans

[0070] Soybeans were soaked in five-fold volumes of water overnight and drained. Ten parts by weight of the soaked soybeans was placed into a pan, mixed with 35 parts by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method of Example A-1, and 35 parts by weight of water. The mixture was boiled over mild heat to produce a concentration of about 70% as determined by a brix apparatus and to obtain boiled beans. The beans were placed into a basket and dried with air heated to 40° C. overnight to product a composition containing soy beans having an active oxygen eliminating activity of about 25 units/g of composition. The product retained the color tint of the boiled soybeans. When tasted, the taste and flavor spread throughout the mouth and stimulated the appetite. In this product, trehalose hydrate is present within the soybeans' tissues, and the hydrate retained the issues. The product was a solid food having satisfactory taste and mouth feel. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals and edible fibers inherent in the soybeans, and augments the active oxygen eliminating activity to living bodies. Thus, the composition can be used to maintain and promote health, prevent aging and geriatric disease, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-17 Composition Containing Apple

[0071] Fresh apples were peeled and sliced to produce slices about 5 mm in thickness, which were then soaked for 30 minutes in an about 75% solution prepared by dissolving a reduction inhibitory agent for active oxygen eliminating activity obtained by the method in Example A-1 in water by heating at 80° C. The apples and solution were placed into a basked and dried at 35° C. overnight to obtain a composition containing apples having an active oxygen eliminating activity of about 50 units/g composition. The production retained the color tint of apples. When tasted, the taste and flavor spread throughout the mouth and stimulated the appetite. In this product, trehalose hydrate is present within the apples' tissues, and the hydrate retained the tissues. The product was a solid food having satisfactory taste and mouth feel. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals and edible fibers inherent in the apples, and augments the active oxygen eliminating activity to living bodies. Thus, the composition can be used to maintain and promote health, prevent aging and geriatric disease, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-18 Composition Containing Carrot

[0072] Fresh carrots were sliced and five parts by weight of the sliced carrots was placed in an about 73% syrup which was mildly boiling at about 105° C. and which had been prepared by dissolving by heating 140 parts by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method in Example A-1 in 60 parts by weight of water. The resulting mixture was concentrated by boiling up to a temperature of about 110° C. for 20 minutes and was dehydrated by heating to produce a fried-like product. Thereafter, the product was placed into a basket, dried by air heated to 60° C. for five hours to obtain a composition containing sliced carrots having an active oxygen eliminating activity of about 200 units/g composition. The composition was a fried-like product of carrot, to which crystalline trehalose hydrate adhered. It retained the inherent color to carrot and stimulated the appetite. The product had a relatively low hygroscopicity, satisfactory shelf life, and desirable handle ability. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals and edible fibers inherent in the carrots, and augments the active oxygen eliminating activity to living bodies. Thus, the composition can be used to maintain and promote health, prevent aging and geriatric disease, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-19 Composition Containing Apple

[0073] Fresh apples were peeled and sliced to a thickness of about 5 mm. Four parts by weight of the sliced apples was then placed into an about 80% syrup heated to about 110° C., which had been prepared by dissolving by heating in water 140 parts by weight of a reduction inhibitory agent for active oxygen eliminating activity obtained by the method of Example A-1 and concentrated by boiling at a temperature of about 114° C. for about 10 minutes and dehydrated by heating to produce a fried-like product. Thereafter, the product was placed into a basket, dried by air heated to 35° C. overnight to product a composition containing sliced applied having an active oxygen eliminating activity of about 30 units/g composition. The composition was a fried-like apple product, to which crystalline trehalose hydrate adhered. It retained the inherent color of apple and stimulated the appetite. The product had a relatively low hydroscopicity, satisfactory shelf life, and desirable handleabiliy. Intake of the composition enriches living bodies with functional ingredients such as vitamins, minerals and edible fibers inherent in the soybeans, and augments the active oxygen eliminating activity to living bodies. Thus, the composition can be used to maintain and promote health, prevent aging and geriatric disease, promote the treatment of incurable diseases, and prevent carcinogenesis. Thus, the composition can be used as a food product, cosmetic, pharmaceutical, or components or processing intermediates thereof.

EXAMPLE B-20 Chewing Gum

[0074] Three parts by weight of a gum base was heated to be softened, then mixed with six parts by weight of crystalline trehalose hydrate, one part by weight of a composition containing aojiso obtained by the method of Example B-9, and 0.01 parts by weight of a tea extract. The resulting mixture was admixed with adequate amounts of flavor and color, kneaded by a roll in a conventional manner, shaped, and packaged into a product having an active oxygen eliminating activity of over 120 units/g product. The product is a chewing gum with satisfactory texture, flavor, and taste. The product containing trehalose as a saccharide is not easily assimilated by dental caries inducing microorganisms, and it does not cause dental caries.

EXAMPLE B-21 Dango (a Rice Paste)

[0075] Ten parts by weight of glutinous rice starch was mixed with 12 parts by weight of water, and the mixture was gelatinized by heating, then kneaded with 0.5 part by weight of a composition containing mugwort obtained by the method of Example B-10. The mixture thus obtained was shaped and packaged in a conventional manner to produce a dango having an active oxygen eliminating activity of over 12 units/g content. The product is a yomogi-dango, i.e., a rice paste with mugwort, having a satisfactory color tint of yomogi, flavor, taste, and mouth feel.

EXAMPLE B-22 Nutritional Product

[0076] A composition was prepared from the following ingredients: 10 parts by weight of crystalline α-maltose, 1.1 parts by weight of glycine, 0.18 part by weight of sodium glutamate, 1.2 parts by weight of salt, one part by weight of citric acid, 0.4 part by weight of calcium lactate, 0.3 part by weight of a composition containing spinach obtained by the method of Example B-1, 0.01 part by weight of thiamine, and 0.01 part by weight of riboflavin. Twenty four gram aliquots of the composition were injected into small laminated aluminum bags and heat sealed to obtain a nutritional product for intubtaion having an active oxygen eliminating activity of over 12 units/g composition. One bag of the nutritional product is dissolved in about 300-500 ml of water, and the solution can be administered to subjects orally or through the subjects' nasal cavities, stomachs, or intestines as a parenteral liquid nutritional supplement.

EXAMPLE B-23 Cosmetic Cream

[0077] Two parts by weight of polyoxyethylene glycol monostearate, 5 parts by weight of glyceryl monostearate, self-emulsifying, 2.8 parts by weight of a composition containing aloe obtained by the method in Example B-12, 0.2 part by weight of α-glucosyl rutin, one part by weight of liquid petrolatum, 10 parts by weight of glyceryl tri-2-ethylhexanoate, and an adequate amount of an antiseptic were dissolved by heating in a conventional manner. The resulting solution was admixed with two parts by weight of L-lactic acid, five parts by weight of 1,3-butylene glycol and 66 parts by weight of refined water, and the mixture was emulsified by a homogenizer and admixed with an adequate amount of a flavor under stirring to obtain a cosmetic cream having an active oxygen eliminating activity of over 46 units/g product. The product can be used as a therapeutic or preventive agent for sunburned skin, as a skin beautifying agent, a skin whitening agent, and an agent for inhibiting aging of skin including chloasmas, freckles, pigmentation, and wrinkles.

EXAMPLE B-24 Bath Salts

[0078] Bath salts having an active oxygen eliminating activity of over 220 units/g product were prepared by mixing 26 parts by weight of refined water and adequate amounts of a coloring agent and flavoring agent with 21 parts by weight of sodium DL-lactate, eight parts by weight of sodium pyruvate, five parts by weight of a composition containing citron obtained by the method of Example B-7, one part by weight of α-glucosyl rutin, and 40 parts by weight of ethanol. The product can be suitable used as a skin beautifying agent and skin whitening agent by diluting 100-10,000 fold in hot water in a bathtub. Similarly as above, the product can be used by diluting it in water for a face wash or beauty wash.

EXAMPLE B-25 Ointment

[0079] One part by weight of sodium acetate trihydrate and four parts by weight of calcium DL-lactate were mixed to homogeneity with 10 parts by weight of glycerin, and this mixture was added to a mixture consisting of 50 parts by weight of petrolatum, 10 parts by weight of vegetable wax, five parts by weight of lanolin, 14.5 parts by weight of sesame oil, six parts by weight of a composition containing dokudami obtained by the method of Example B-11, and 0.5 part by weight of peppermint oil. The mixture thus obtained was further mixed to homogeneity into an ointment having an active oxygen eliminating activity of about 54 units/g. The product can be used as an antipyogenic agent, skin beautifying agent, skin whitening agent, and agent for promoting the treatment of skin traumas and burns.

EXAMPLE B-26 Tablet

[0080] Twenty parts by weight of ascorbic acid were mixed to homogeneity with 13 parts by weight of crystalline β-maltose, four parts by weight of corn starch, three parts by weight or a composition containing plantain and trehalose obtained by the method of Experiment 5 according to the present invention. The resulting mixture was formed into a tablet using a 20R punch, 12 mm in diameter. The tablet had an oxygen eliminating activity of over 560 units/g, and can be administered orally to subjects in the amount of 1-10 tablets/adult/day. These tablets can particularly be used for reducing fever and maintaining and controlling stomach and intestinal conditions.

[0081] As described above, the present invention provides a reduction inhibitory agent for active oxygen eliminating activity which comprises trehalose as an effective ingredient, a method for inhibiting the reduction of active oxygen eliminating activity comprising incorporating either trehalose or the reduction inhibitory agent into products to be treated, and a composition which contains plant edible substance and/or plant antioxidant in which the reduction of active oxygen eliminating activity is inhibited by the above method. Intake of the present composition easily nutritionally supplements living bodies and enriches living bodies with functional ingredients such as vitamins, minerals, and edible fibers of plant edible substances. Therefore, the present composition contributes greatly to maintain and promote health, prevent aging and geriatric diseases promote the treatment of incurable diseases, and inhibit carcinogenesis. Thus, the present invention provides a novel health resource as a fourth functional ingredient which can be used in processing and use of plant edible substances. The present invention provides a great contribution to a wide variety of fields, particularly the fields of food products, cosmetics, and pharmaceuticals.

[0082] While there has been described what is at present considered to be the preferred embodiments of the invention, it will be understood that various modifications may be made thereto, and it is intended to cover in the appended claims all such modifications as fall within the true spirit and scope of the invention. 

What is claimed is:
 1. A reduction inhibitory agent for active oxygen eliminating activity comprising an effective amount of trehalose as an active ingredient.
 2. The inhibitory agent according to claim 1 which further contains a saccharide other than trehalose.
 3. The inhibitory agent according to claim 1 which further contains pullulan, cyclodextrin, or mixtures thereof.
 4. The inhibitory agent according to claim 1 in which the trehalose is present in an amount of at least 20 w/w % of said agent, on a dry solid basis.
 5. A composition obtained by incorporating the inhibitory agent of claim 1 into a plant substance with active oxygen eliminating activity to inhibit the reduction of said activity.
 6. The composition according to claim 5 wherein said plant substance is a plant edible substance, a plant antioxidant, or mixtures thereof.
 7. The composition according to claim 6 wherein said plant edible substance is at least one of an organ and a tissue of an edible plant, or a sliced, disrupted, pulverized, dried, pickled, or extracted edible part of said plant.
 8. The composition according to claim 6 wherein said plant antioxidant is a member of the group consisting of plant enzymes, pigments, polyphenols, and vitamins.
 9. The composition according to claim 5 which contains at least about 1 w/w %, on a dry solid basis, of trehalose as an active ingredient.
 10. The composition according to claim 5 wherein the trehalose is present in an amorphous or crystalline hydrous form.
 11. The composition according to claim 5 which contains at least one member selected from the group consisting of essential minerals, edible fibers, saccharides which promote the growth of bifid bacteria, vitamins, and biologically active substances.
 12. The composition according to claim 5 which has been sterilized or dried or sterilized and dried.
 13. The composition according to claim 5 wherein has an active oxygen eliminating activity of at least five units/g or compositions, on a dry solid basis, as determined by the nitroblue tetrazolium test.
 14. The composition according to claim 5 which is in the form of a food product, cosmetic, pharmaceutical, or components or intermediates thereof.
 15. A composition comprising the composition of claim 5 and a food, cosmetic, or pharmaceutical.
 16. A composition comprising at least about 30 w/w % of trehalose on a dry solid basis, and edible plant tissue.
 17. The composition according to claim 16 wherein said edible plant tissue is selected from the group consisting of organs and tissues of an edible plant, and sliced and picked products thereof and the trehalose is in an amorphous or crystalline hydrous form.
 18. The composition according to claim 16 which has been sterilized, dried, or sterilized and dried.
 19. The composition according to claim 16 which is in the form of a solid food product.
 20. The composition according to claim 19 wherein said solid food product has satisfactory mouth feel. 